Abstract: A method for designing circular permuted proteins using two engineered Mu transposons for easy construction of random circular permuted proteins, the two designed transposons being MuCP-ISC (Mu Circular permutation transposon with Integrated Start Codon) and MuCP-ISSC (Mu Circular permutation transposon with Integrated Start and Stop Codon).

Abstract: A method for designing circular permuted proteins using two engineered Mu transposons for easy construction of random circular permuted proteins, the two designed transposons being MuCP-ISC (Mu Circular permutation transposon with Integrated Start Codon) and MuCP-ISSC (Mu Circular permutation transposon with Integrated Start and Stop Codon).

Abstract: A conformation-switching fluorescent protein probe for detection of alpha synuclein oligomers using an alpha synuclein (?S) variant, PG65 (SEQ ID NO: 4), together with a conformation-sensitive fluorescent molecule to create a molecular probe for rapid, specific, and quantitative detection of ?S oligomers.

Abstract: Methods for facile construction of a random domain insertion library (1) with optimal control of composition and length of inter-domain linker residues and (2) mediated by sticky-end ligation between host and guest DNA fragments. To develop such a method, we engineered a Mu transposon. The method exploits transposition of the engineered Mu transposon, which, upon removal, allows for sticky-end ligation between host and guest DNA fragments. We used a gene coding for xylanase from bacillus circulans (BCX) as a guest DNA sequence and the plasmid PUC19 containing lacZ? as the target for insertion (i.e., a host DNA sequence). Results demonstrate that the method enables facile construction of a random domain insertion library with optimal control of composition and length of inter-domain linker residues.

Abstract: A conformation-switching fluorescent protein probe for detection of alpha synuclein oligomers. The use of intrinsically disordered proteins as conformation-switching biosensors. The use of an alpha synuclein (?S) variant, PG65, together with conformation-sensitive fluorescence to create a molecular probe for rapid, specific and quantitative detection of ?S oligomers.

Abstract: Methods for facile construction of a random domain insertion library (1) with optimal control of composition and length of inter-domain linker residues and (2) mediated by sticky-end ligation between host and guest DNA fragments. To develop such a method, we engineered a Mu transposon. The method exploits transposition of the engineered Mu transposon, which, upon removal, allows for sticky-end ligation between host and guest DNA fragments. We used a gene coding for xylanase from bacillus circulans (BCX) as a guest DNA sequence and the plasmid PUC19 containing lacZ? as the target for insertion (i.e., a host DNA sequence). Results demonstrate that the method enables facile construction of a random domain insertion library with optimal control of composition and length of inter-domain linker residues.

Abstract: A peptide probe that generates fluorescence signals rapidly upon recognition of various A? aggregates without significant perturbation of samples. The present peptide probes display an increase in fluorescence signals upon coincubation with A? oligomers, but neither monomeric/dimeric species nor fibrils. The detection can occur within an hour or two without any additional sample preparation and incubation steps.

Abstract: A strategy to improve protein stability by domain insertion. TEM 1 beta-lactamase (BLA) and exo-inulinase, as model target enzymes, are inserted into a hyperthermophilic maltose binding protein from Pyrococcus furiosus (PfMBP). Unlike conventional protein stabilization methods that employ mutations and recombinations, the inventive approach does not require any modification on a target protein except for its connection with a hyperthermophilic protein scaffold. For that reason, target protein substrate specificity was largely maintained, which is often modified through conventional protein stabilization methods. The insertion was achieved through gene fusion by recombinant DNA techniques.

Abstract: A peptide probe that generates fluorescence signals rapidly upon recognition of various A? aggregates without significant perturbation of samples. The present peptide probes display an increase in fluorescence signals upon coincubation with A? oligomers, but neither monomeric/dimeric species nor fibrils. The detection can occur within an hour or two without any additional sample preparation and incubation steps.

Abstract: A strategy to improve protein stability by domain insertion. TEM 1 beta-lactamase (BLA) and exo-inulinase, as model target enzymes, are inserted into a hyperthermophilic maltose binding protein from Pyrococcus furiosus (PfMBP). Unlike conventional protein stabilization methods that employ mutations and recombinations, the inventive approach does not require any modification on a target protein except for its connection with a hyperthermophilic protein scaffold. For that reason, target protein substrate specificity was largely maintained, which is often modified through conventional protein stabilization methods.